Figure 2. Denosumab significantly reduces mammosphere formation in BRCA1^mut/+ breast epithelial cells.

(a) Mammosphere-forming efficiency (MSFE) of BRCA1^+/+ and BRCA1^mut/+ MCF10A isogenic cell pairs in the absence or presence of 100 μg/mL denosumab was calculated after 7 days and expressed as a percentage (mean ± SD); three technical replicates per n; n = 3 biological replicates. The MSFE of vehicle-alone control cells was normalized to one in each isogenic model. Re-feeding of mammosphere cultures with denosumab and/or sphere medium was performed on day 4. *p<0.05. (b) Monolayers of MCF10A BRCA1^+/+ and BRCA1^mut/+ cells were pre-treated with 100 μg/mL denosumab for 3 days, trypsinized and re-plated for mammosphere assays in the absence of denosumab. The MSFE of vehicle-alone control cells was normalized to one; three technical replicates per n; n = 3 biological replicates. *p<0.05.